Ultra-high performance liquid chromatography tandem mass spectrometry method for detection of α-hydroxybutyrate

Objective To establish an ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) method for detecting α-hydroxybutyrate (α-HB) in serum. Methods Electrospray ionization negative ion and multiple reaction monitoring mode were used to detect serum α-HB. The linearity, low limits of quantification, precision, recovery and interference of UHPLC-MS/MS were evaluated. The reference interval of this method was established in 130 serum samples (62 males and 68 females) from Shanghai East Hospital. Dixon method was used to judge the outliers and K-S test was used to analyze the data normality. The standard curve was scored by linear regression analysis. Results The total run time was 4 min of UHPLC-MS/MS method for the determination of α-HB. It has a good linear relationship in the range of 0.5-40.0 mg/L(r=0.999 4); the low limit of quantification was 0.5 mg/L; the in-batch and inter-batch coefficient of variation precision were less than 4.1% and 6.3%, respectively; the recovery ranged between 95.8% and 103.8%. Hemolytic samples (about 5 g/L hemoglobin), lipemic samples (about 12 mmol/L triglyceride), icteric samples (about 150 μmol/L total bilirubin) had no significant interference to the detection. The reference range of the apparent healthy population was 1.46-6.48 mg/L. Conclusions A method for the determination of serum α-HB by UHPLC-MS/MS was established. The method was simple, rapid, and could be used for the detection of clinical samples. Key words: Butyric Acids; Chromatography, High Pressure Liquid; Insulin resistance